Isolation of an in Vitro Affinity-Matured, Thermostable ``Headless'' HA Stem Fragment That Binds Broadly Neutralizing Antibodies with High Affinity

Najar, Tariq Ahmad and Kar, Uddipan and Flynn, Jessica A and Varadarajan, Raghavan (2018) Isolation of an in Vitro Affinity-Matured, Thermostable ``Headless'' HA Stem Fragment That Binds Broadly Neutralizing Antibodies with High Affinity. In: BIOCHEMISTRY, 57 (26). pp. 3817-3829.

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Abstract

The surface glycoprotein hemagglutinin (HA) of influenza virus is the primary target for the design of an effective universal influenza vaccine as it is capable of eliciting broadly cross-reactive antibodies against different HA subtypes. Several monoclonal antibodies targeting the stem region of HA that are able to neutralize various subtypes of influenza virus have been isolated in the recent past. Designing a stable, HA stem immunogen that attains a native-like conformation and can elicit such antibodies has been a challenge. We describe the affinity maturation of a previously designed stem immunogen (H1HA6) by random mutagenesis, followed by selection using yeast surface display. The affinity-matured mutant protein (H1HA6P2), upon bacterial expression, attained a stable, native-like, trimeric conformation without any heterologous trimerization motif and showed a significant improvement in thermal stability and binding to several stem specific, conformation-sensitive, broadly neutralizing antibodies (bnAbs) relative to H1HA6. These results point to an effective strategy for the design of stabilized HA stem immunogens that can be tested for their protective ability.

Item Type:	Journal Article
Publication:	BIOCHEMISTRY
Publisher:	AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036 USA
Additional Information:	Copyright of this article belong to AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036 USA
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	01 Aug 2018 15:22
Last Modified:	01 Aug 2018 15:22
URI:	http://eprints.iisc.ac.in/id/eprint/60336

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